Greg, I can help you with the last question. Your human AB serum probably has complement activity which will definitely lyse any antibody labelled cells. Heat de-activation will usually take care of this. But I would recommend using serum from the host animal in which the antibody was raised-mouse or goat. It should more effectively block any non-specific binding. Art On Thu, 11 Mar 1999, Greg Neely wrote: > > > 5. I just tried to stain my cells for surface markers in PBS, 0.1% > sodium azide, but 1% human AB serum instead of fetal calf serum, hoping > this might better block non-specific interactions. Instead my cell > count was absurdly low, and there was tons of dead cells. Has anyone > seen this? Is this why one does not traditionally wash/stain cells in > human serum? > > > Art Roberts Dept. of Medicine Robert Wood Johnson Medical School email: robertar@umdnj.edu phone: 732-235-7790 Fax: 732-235-7792
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